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  • 1
    Electronic Resource
    Electronic Resource
    Studies on the preparation and on the properties of sea snail enzymes (1984)
    Springer
    Hydrobiologia 116-117 (1984), S. 319-320 
    Details
    ISSN: 1573-5117
    Keywords: seaweed ; enzyme ; protoplasts ; decomposition ; preparation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027694
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  • 2
    Electronic Resource
    Electronic Resource
    Flow cytometric analysis of tobacco and cowpea protoplasts infected in vivo and in vitro with alfalfa mosaic virus (1983)
    Springer
    Plant molecular biology 2 (1983), S. 19-25 
    Details
    ISSN: 1573-5028
    Keywords: alfalfa mosaic virus ; protoplasts ; viral antigen detection flow cytometry ; fluorescence-activated cell sorter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunofluorescence flow cytometry was used to study the distribution of viral antigen in protoplast populations. Protoplasts were isolated from healthy and alfalfa mosaic virus (AMV) infected tobacco leaves (designated in vivo infected). Furthermore isolated tobacco and cowpea protoplasts were infected in vitro with AMV. The FITC-conjugated antibodies could penetrate formaldehyde fixed protoplasts. The flow cytometric measurements were rapid and reproducible. Comparable immunofluorescence patterns were found for all infected samples (per sample 104 protoplasts were measured). Infectious virus could only be detected in in vivo infected tobacco protoplasts and in in vitro infected cowpea protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00187572
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  • 3
    Electronic Resource
    Electronic Resource
    An alfalfa mosaic virus RNA 2 mutant, which does not induce a hypersensitive reaction in cowpea plants, is multiplied to a high concentration in cowpea protoplasts (1983)
    Springer
    Plant molecular biology 2 (1983), S. 85-88 
    Details
    ISSN: 1573-5028
    Keywords: alfalfa mosaic virus ; mutant ; protoplasts ; supplementation ; repression ; host response ; hypersensitive reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A mutant of alfalfa mosaic virus (AMV), which in contrast to wild type (wt) can invade cowpea plants systemically, is replicated more efficiently in cowpea protoplasts than the wt. Mutant preparations isolated from infected cowpea protoplasts contained a higher amount of middle component (M, containing RNA 2) than wt preparations. Both in cowpea plants and in cowpea protoplasts a wt phenotype is obtained upon addition of wt M to this mutant, suggesting a correlation between the type of plant reaction evoked by the virus infection and the regulation of viral RNA synthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01595169
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Improved protoplast culture and agarose media (1983)
    Springer
    Plant cell, tissue and organ culture 2 (1983), S. 217-226 
    Details
    ISSN: 1573-5044
    Keywords: protoplasts ; agarose ; Nicotiana tabacum ; Nicotiana plumbaginifolia ; Daucus carota ; Hyoscyamus muticus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Media solidified with agarose resulted in higher plating efficiency of protoplasts than commonly used agar media. Improved culture efficiency was observed with mesophyll protoplasts of Nicotiana tabacum and N. plumbaginifolia and with protoplasts isolated from cell lines of Daucus carota, Hyoscyamus muticus and two lines of N. tabacum. The improvement with agarose was consistent over a wide cell density range and also for different media. The positive effect was not due to the lower temperature at which the protoplasts could be plated. Culture experiments with mixtures of different agar types generally gave intermediate division frequencies. There was no obvious effect on plating efficiency in experiments where diffusion was permitted between agar and other agar types.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033560
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  • 5
    Electronic Resource
    Electronic Resource
    Rotation of cells in an alternating electric field theory and experimental proof (1982)
    Springer
    The journal of membrane biology 67 (1982), S. 13-26 
    Details
    ISSN: 1432-1424
    Keywords: cell rotation ; polarization ; protoplasts ; dielectric breakdown
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Protoplasts ofAvena sativa rotate in an alternating electric field provided that at least two cells are located close to each other. An optimum frequency range (20 to 30 kHz) exists where rotation of all cells exposed to the field is observed. Below and above this frequency range, rotation of some cells is only occasionally observed. The angular velocity of rotation depends on the square of the electric field strength. At field strengths above the value leading to electrical breakdown of the cell membrane, rotation is no longer observed due to deterioration of the cells. The absolute value of the angular velocity of rotation at a given field strength depends on the arrangement of the cells in the electric field. A maximum value is obtained if the angle between the field direction and the line connecting the two cells is 45o. With increasing distance between the two cells the rotation speed decreases. Furthermore, if two cells of different radii are positioned close to each other the cell with the smaller radius will rotate with a higher speed than the larger one. Rotation of cells in an alternating electric field is described theoretically by interaction between induced dipoles is adjacent cells. The optimum frequency range for rotation is related to the relaxation of the polarization process in the cell. The quadratic dependence of the angular velocity of rotation on the field strength results from the fact that the torque is the product of the external field and the induced dipole moment which is itself proportional to the external field. The theoretical and experimental results may be relevant for cyclosis (rotational streaming of cytoplasm) in living cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01868644
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  • 6
    Electronic Resource
    Electronic Resource
    Sugarcane tissue and protoplast culture (1981)
    Springer
    Plant cell, tissue and organ culture 1 (1981), S. 149-164 
    Details
    ISSN: 1573-5044
    Keywords: Saccharum ; cell culture ; protoplasts ; embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Experiments are described which improve the protocols for initiating in vitro cultures of sugarcane and allowing efficient regeneration of plants even after 30 months of callus proliferation. Procedures adopted included use of leaf base explants, CS medium with 3 mg/l 2, 4-D and 0.25 mg/l kinetin for callus initiation and growth, MS medium with 0.5 mg/l IAA and 1 mg/l BAP for shoots, MS medium with 5 mg/l NAA and 7% (wt/vol) sucrose for rooting of shoots. Casein hydrolysate (N-Z amine) significantly shortened the lag period in the growth of sugarcane suspension cultures, but did not increase the rate of growth following the lag phase. Protoplasts isolated from two types of cultures could be grown to re-establish cell cultures but no plants have yet been regenerated derived from isolated protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02318913
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  • 7
    Electronic Resource
    Electronic Resource
    Winged-bean protoplasts: Isolation and culture to callus (1981)
    Springer
    Plant cell, tissue and organ culture 1 (1981), S. 201-209 
    Details
    ISSN: 1573-5044
    Keywords: protoplasts ; suspension cells ; callus formation ; winged bean ; Psophocarpus tetragonolobus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A system was developed for protoplast isolation and culture from suspension cultured cells of winged bean,Psophocarpus tetragonolobus. Cells from a three-day-old suspension were incubated in an enzyme mixture containing 6% cullulysin, 1% Macerase, 1% desalted Rhozyme, 0.4M sorbitol, and 0.1M CaCl2 at pH 5.5. Average yields of protoplasts were 6.5 × 106 per gram fresh weight of cells. Protoplasts were cultured in modified B5 medium containing 68.4 g/l glucose, 250 mg/l xylose, 0.1 mg/l 2,4-D, 0.5 mg/l BAP, 250 mg/l N-Z amine type AS, and 20 ml/l coconut water. After 24 h of culture, the protoplasts had synthesized a new wall, and in three days had begun division. The optimum plating density was 1–2 × 103 protoplasts/ml. The division frequency ranged between 40%–60% for most experiments with a high of 72% in one experiment. After three weeks, cell colonies could be transferred to solid MS medium containing N-Z amine and coconut water where callus developed. This protoplast system is technically comparable to soybean for experiments concerned with genetic manipulation involving legumes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02318916
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  • 8
    Electronic Resource
    Electronic Resource
    factors increasing the fusion efficiency ofBacillus subtilis (1980)
    Springer
    Bulletin of experimental biology and medicine 89 (1980), S. 666-668 
    Details
    ISSN: 1573-8221
    Keywords: protoplasts ; fusion of protoplasts ; regeneration ; “primary prototrophs.”
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00835815
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  • 9
    Electronic Resource
    Electronic Resource
    Protoplast technology for the breeding of top-fruit trees (Prunus, Pyrus, Malus, Rubus) and woody ornamentals (1955)
    Springer
    Euphytica 85 (1955), S. 287-294 
    Details
    ISSN: 1573-5060
    Keywords: protoplasts ; protoclonal variation ; somatic hybridization ; top-fruit trees ; woody ornamentals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Until recently, temperate fruit trees and woody ornamentals have been regarded as recalcitrant to biotechnological breeding approaches based on protoplasts. This however should no longer be the case, as procedures are now available, not only for the regeneration of complete plants from protoplasts of various tissues of such species, but also for the exploitation of protoplast technology for their genetic manipulation. This paper will examine the recent advances and state of the art in this domain, with particular attention to the use of protoplast technology as a novel tool in the breeding of rosaceous top-fruit tree species and woody ornamentals. Problems and their solutions within the context of regenerating plants from isolated protoplasts of stone (Prunus spp.), pome (Pyrus spp., Malus spp.) and small (Rubus spp.) fruits, and of several shrubby ornamental genotypes (Lonicera spp., Weigela spp., Forsythia spp., Cotoneaster spp.) will be addressed. Interspecific (Prunus spinosa + Prunus cerasifera) and intergeneric (Forsythia spp. + Syringa spp.) somatic hybridization within this group of species, as well as the use of protoplasts for host/pathogen interaction studies (Pyrus/Erwinia amylovora) will also be discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023958
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  • 10
    Electronic Resource
    Electronic Resource
    Gene specificity is maintained in transient expression assays with protoplasts derived from different tissues of barley (1955)
    Springer
    Euphytica 85 (1955), S. 203-207 
    Details
    ISSN: 1573-5060
    Keywords: barley ; electroporation ; PEG-mediated DNA uptake ; promoter analysis ; protoplasts ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In some cereal species that are still recalcitrant to stable transformation and regeneration, transient expression in isolated protoplasts is a useful tool for the study of gene expression and regulation. We have successfully applied these techniques to barley protoplasts derived from developing endosperm, aleurone, leaves and roots in order to characterize functionally cis-acting motives in two gene promoters, corresponding to trypsin inhibitor BTI-CMe and to sucrose synthase Ss1. Gene specificity is maintained in transient expression assays with protoplasts isolated from these different barley tissues and the pattern of expression parallels the mRNA levels observed for the corresponding genes in the same tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023949
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  • 11
    Electronic Resource
    Electronic Resource
    The phenotypic characterisation of R2 generation transgenic rice plants under field and glasshouse conditions (1955)
    Springer
    Euphytica 85 (1955), S. 395-401 
    Details
    ISSN: 1573-5060
    Keywords: Oryza sativa L. cv. Taipei 309 ; rice ; protoplasts ; direct DNA uptake ; kanamycin-resistant transgenic plants ; field trial ; glasshouse trial ; neomycin phosphotransferase II (npt II) gene ; gene expression and inheritance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The phenotypes of seed progeny (R2 generation) of Oryza sativa L. cv. Taipei 309, which carried the neomycin phosphotransferase II (npt II) gene, were compared with those of non-transformed, protoplast-derived plants of the same generation and non-transformed, seed-derived plants under field and glasshouse conditions. Under both conditions the transgenic plants were generally smaller, took longer to flower and had reduced fertility. Significant differences were observed between individuals within the group of transgenic plants. The npt II gene was present in most of the transgenic plants, but NPT II activity was only detected in a minority of individuals.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023972
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  • 12
    Electronic Resource
    Electronic Resource
    Studies of the mechanism of transgene integration into plant protoplasts: improvement of the transformation rate (1955)
    Springer
    Euphytica 85 (1955), S. 53-61 
    Details
    ISSN: 1573-5060
    Keywords: bleomycin ; direct gene transfer ; expression ; irradiation ; petunia ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The production of transgenic plants by means of direct gene transfer to protoplasts is now a widely-used technique. The biological mechanisms underlying the transformation are still poorly understood, but many investigations have attempted to shed light on some components of this process. Varying the experimental conditions has in some cases led to better transformation rates, but further improvements of the protocols are possible. Such improvements will require a better understanding of how the alien DNA enters the cells, becomes integrated into the chromosomes and is treated as a part of the plant genome. Irradiation with sublethal doses of X-rays or UV-light has been shown to increase the transformation frequency, while certain drugs have been shown to act in a similar manner. The effects of these and other factors are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023930
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